Promene u limfatičnim organima komercijalnih pilića nakon vakcinacije protiv Marekove bolesti - histološka i stereološka analiza

The aim of this study was to investigate histomorphometrical characteristics of the thymus, bursa of Fabricius and spleen in the chickens vaccinated with a vaccine against Marek’s disease. For this purpose, we used newly hatched chickens of the light hybrid line, obtained from a local hatchery. The chickens were vaccinated on the 5th day after hatching with a bivalent cell-associated Marek's disease vaccine (PFU-2000 per dose). On day 13 both vaccinated chickens and unvaccinated controls were sacrificed, and thymus, bursa and spleen were removed and processed for light microscopy. The serial tissue sections, hematoxylin-eosine stained, were used for histomorphometric analysis. Vaccination against Marek’s disease decreased the relative mass of the lymphoid organs, and caused significant damage of the thymus and spleen in experimental chicken. In addition, vaccination, similar to Marek's disease virus, induced morphometric changes in the lymphoid organs. Namely, it significantly decreased the diameter and volume of lymphoid follicles, volume of follicular medulla and number of cells in the follicular cortex in the bursa of Fabricius. In the thymus, vaccination reduced the thymus volume and the absolute number of thymocytes. However, vaccination against Marek’s disease caused an increase in the diameter, number and volume of lymphoid follicles in the spleen. The present data suggest that vaccination against Marek’s disease was able to induce the immune response in processed organs, although it reduced the mass and number of lymphocytes in the major lymphoid organs.Cilj ovih ispitivanja su bile histomorfometrijske karakteristike timusa, burze Fabricii i slezine pilića vakcinisanih protiv Marekove bolesti. Za istraživanja su korišćeni jednodnevni pilići lake hibridne linije, gajeni u standardnim uslovima. Pilići su vakcinisani 5. dana od izleganja, bivalentnom vakcinom protiv Marekove bolesti (PFU 2000 po dozi). Trinaestog dana od izleganja kontrolni i vakcinisani pilići su žrtvovani, uzeti su im timus, burza Fabricii i slezina koji su pripremljeni za svetlosno mikroskopsku analizu. Za analizu su korišćeni serijski preseci limfatič nih organa, bojeni metodom hematoksilin-eozin. Dobijeni rezultati su ukazali da vakcinacija protiv Marekove bolesti dovodi do smanjenja relativne mase timusa, burze i slezine, kao i povećanja indeksa oštećenja timusa i slezine. Takođe je ustanovjleno da vakcinacija dovodi do promena morfometrijskih parametara u limfatičnim organima. U burzi Fabricii je uočeno značajno smanjenje dijametra i zapremine limfocitnih folikula, zapremine medule folikula i broja ćelija u korteksu folikula. U timusu vakcinacija dovodi do redukcije zapremine timusa i apsolutnog broja timocita. Međutim, vakcinacija protiv Marekove bolesti u slezini dovodi do povećanja dijametra limfocitnih folikula, kao i do povećanja njihovog broja i zapremine. Rezultati ove studije su ukazali da vakcinacija pilića protiv Marekove bolesti, iako redukuje broj T i B limfocita, indukuje razvoj imunskog odgovora

The Ninth Dimension of National Culture: Unpacking Cross-Cultural Communication Styles

This article aims to study the communication styles among different national cultures, that is, we examine the relationship between national culture, based on Hofstede model, and communication styles. In order to investigate the role of national culture, it is fundamental to first analyse the communication styles and then identify how these are related to each other. With a purpose to identify differences (or similarities) in communication styles across selected countries, a factor analysis was conducted, combined with an ANOVA test. Based on a sample from 10 different cultures: Germany, Sweden, Japan, China, Russia, Italy, the United States, the United Kingdom, Serbia and North Macedonia, and using communication assessment instrument as well as the data on Hofstede’s six dimensions of national culture model, the findings show that different national cultures practise different communication styles. When Professional-Casual communication style is concerned, Germany is the most professional, while Japan is the most casual; that is, the least professional of all, while the analysis of the Cold-Warm communication styles leads us to the fact that Sweden is the coldest, and the US is the warmest country. The study’s significance is hopefully fundamental since it proposes an additional dimension, which is as frequent among cultures as it is rooted deeply in each culture. In this way, the article recommends that the countries ought to both comprehend their national culture and utilise it as a “tool” for understanding other cultures as well

The Ninth Dimension of National Culture: Unpacking Cross-Cultural Communication Styles

This article aims to study the communication styles among different national cultures, that is, we examine the relationship between national culture, based on Hofstede model, and communication styles. In order to investigate the role of national culture, it is fundamental to first analyse the communication styles and then identify how these are related to each other. With a purpose to identify differences (or similarities) in communication styles across selected countries, a factor analysis was conducted, combined with an ANOVA test. Based on a sample from 10 different cultures: Germany, Sweden, Japan, China, Russia, Italy, the United States, the United Kingdom, Serbia and North Macedonia, and using communication assessment instrument as well as the data on Hofstede’s six dimensions of national culture model, the findings show that different national cultures practise different communication styles. When Professional-Casual communication style is concerned, Germany is the most professional, while Japan is the most casual; that is, the least professional of all, while the analysis of the Cold-Warm communication styles leads us to the fact that Sweden is the coldest, and the US is the warmest country. The study’s significance is hopefully fundamental since it proposes an additional dimension, which is as frequent among cultures as it is rooted deeply in each culture. In this way, the article recommends that the countries ought to both comprehend their national culture and utilise it as a “tool” for understanding other cultures as well

Diferentovanje timocita u organ kulturi timusa odraslih pacova

To investigate the differences between thymocytes development in vivo and in vitro, thymus lobe fragments from 12-weeks old male Albino Oxford rats were cultivated over a 7-days period. In the controls and cultivated thymic lobes fragments were evaluated and the viability, apoptosis and cell cycle of thymocytes, as well as the histological characteristics of thymic tissue. Additionally, we analyzed the expression of CD4, CD8 and TCRαβ on thymocytes by flow cytometry. The obtained results showed that thymus cellularity decreased during cultivated time due to expanded apoptosis, decreased proliferation and the absence of progenitors reseeding thymus. The relative proportion of thymocyte subsets in the first 24 hours of culture remained similar as in the control. However, cultivation for 3 and 7 days modulated the relative proportions between thymoctye subsets. The percentage of DP TCRαβlow increased, DP TCRαβhi subset remained unchanged, both SP TCRαβhi subsets decreased while the same mature SP phenotype dominated in culture media. These results demonstrate that cultivated thymic fragments retain the capacity for T cell development, although cultivation modulates this process.Sa namerom da ispitamo razlike između in vivo i in vitro sazrevanja timocita gajili smo fragmente lobusa timusa poreklom od mužjaka Albino Oksford pacova, starih dvanaest nedelja, u vremenskom periodu od sedam dana. Nakon kultivacije određivani su vijabilnost, apoptoza i ćelijski ciklus timocita, kao i histološke osobine timusnog tkiva. Takođe je analizirano ispoljavanje markera diferentovanja CD4, CD8 and TCRαβ na površini timocita metodom tečne citofluorometrije. Dobijeni rezultati su ukazali da sedmodnevna kultivacija dovodi do smanjenja broja ćelija u timusu usled povećane apoptoze, smanjene proliferacije i odsustva ulaska progenitora timocita. Tokom prvih 24 sata kultivacije ne dolazi do promena u odnosima timocitnih populacija. Međutim, duže vreme kultivacije - 3 i 7 dana moduliše relativne odnose između timocitnih subpopulacija - povećava se procenat DP TCRαβlow, procenat DP TCRαβhi timocita ostaje nepromenjen, dok su procenti ćelija oba subseta SP TCRαβhi smanjeni, mada je prisustvo pomenutih SP subsetova dominantno u medijumu za kultivaciju. Navedeni rezultati pokazuju da kultivisani fragmenti timusnog tkiva zadržavaju sposobnost da podrže sazrevanje timocita u jednostruko pozitivne T ćelije, mada je diferentovanje timocita donekle modulisano kultivacijom

Optimizacija metode za izolaciju epitelnih stanica iz nežljezdanog dijela želuca štakora za protočnu citometriju

Traditional methods in cell proliferation studies are based on immunohistochemical detection of proliferating cells in the target tissue. Since they are time consuming, optimization of novel, more efficient methods is important for large scale proliferation studies. In this study, we aimed to optimize the isolation of single epithelial rat forestomach cells for flow cytometry. As a marker of cellular proliferation we used the Ki-67 antibody to detect this nuclear protein expressed in proliferating cells. We also performed immunohistochemical detection of Ki-67 positive cells and propidium iodide staining to validate the results. 3-tert- butyl -4-hydroxyanisole was used as the positive control to ensure cellular proliferation. The results showed that isolation of epithelial cells with collagenase, trypsin and cell strainer ensures great cell viability (>95%) and the purity of the samples. Flow cytometry and immunostaining with the Ki-67 antibody indicated that 3-tert- butyl-4-hydroxyanisole treatment leads to a significant increase in proliferation. A significant positive correlation was observed between the results obtained by immunohistochemistry and flow cytometry, but the flow cytometric data had a smaller measurement error, suggesting the equal sensitivity and greater accuracy of this method. Propidium iodide staining showed that the percentage of cells in the G2+S phase of the cell cycle correlated positively with the percentage of Ki-67 positive cells assessed by flow cytometry, indicating that Ki-67 positive cells reflect an active dividing cell pool. We conclude that the isolation of forestomach epithelial cells described is a simple and reliable method for obtaining viable cells for use in flow cytometry. Compared to immunohistochemistry, flow cytometric detection of the Ki-67 antigen is equally sensitive, but much faster and provides more accurate results.Tradicionalne metode u ispitivanju stanične proliferacije temelje se na imunohistokemijskom otkrivanju proliferacijskih stanica u ciljanom tkivu. Kako su dugotrajne, optimizacija novih i učinkovitijih metoda važna je za velika istraživanja o proliferaciji. U ovom smo radu željeli optimizirati izolaciju epitelnih stanica prednjeg želuca štakora za protočnu citometriju. Kao marker stanične proliferacije koristili smo Ki-67 protutijelo za otkrivanje ovoga nuklearnog proteina izraženog u proliferacijskim stanicama. Također smo učinili imunohistokemijsku detekciju Ki- 67 pozitivnih stanica i bojenje propidij-jodidom kako bismo potvrdili rezultate. Butil-hidroksianizol korišten je kao pozitivna kontrola da se osigura stanična proliferacija. Rezultati su pokazali da izolacija epitelnih stanica s kolagenazom, tripsinom i staničnim cjedilom osigurava veliku vijabilnost stanica (> 95 %) i čistoću uzoraka. Protočna citometrija i Ki-67 bojenje pokazali su da tretman butil-hidroksianizolom dovodi do znakovitog porasta proliferacije. Primijećena je znakovita pozitivna korelacija između rezultata dobivenih imunohistokemijom i protočnom citometrijom, dok su protočni citometrijski podaci imali manju pogrešku mjerenja, što upućuje na jednaku osjetljivost i veću točnost ove metode. Bojenje propidij-jodidom pokazalo je da postotak stanica u G2+S fazi staničnog ciklusa pozitivno korelira s postotkom Ki-67 pozitivnih stanica procijenjenih protočnom citometrijom, što upućuje na to da Ki-67 oslikava stanice u aktivnoj diobi. Zaključujemo da je opisana izolacija epitelnih stanica prednjeg želuca štakora jednostavna i pouzdana metoda za dobivanje održivih stanica za upotrebu u protočnoj citometriji. U usporedbi s imunohistokemijom, protočna citometrijska detekcija antigena Ki-67 jednako je osjetljiva, ali mnogo brža i daje točnije rezultate

Somatostatin-14 izaziva različite promene u timusima prepubertalnih i mladih odraslih pacova

Bearing in mind the role of somatostatin in thymus functions, and changes of somatostatin level and expression of its receptors during postnatal life, the aim of this study was to investigate whether centrally applied SRIH-14 induces different changes in the thymic compartments and thymocyte profile in peripubertal and young adult rats. To this end, 4- and 10-week-old male AO rats were cannulated and treated intracerebroventriculary with three doses of SRIH-14, applied every other day. In peripubertal rats, SRIH-14 decreases thymic relative weight and volume, as well as the volume of thymic compartments, especially of deep cortex, as a result of thymocytes loss by apoptosis. Also, SRIH-14 increases the percentage of immature thymocytes preceding the DPTCRαβlow cells (DNTCRαβ-/low, DPTCRαβ-, SPCD8TCRαβ-/low and SPCD4TCRαβ-/low), decreases the percentages of DPTCRαβlow and DPTCRαβhi cells, while the relative proportion of CD4+/CD8+TCRαβhi cells remained unaltered. In young adult rats, SRIH-14 does not lead to changes in relative thymus weight, although decreases the thymic cortex cellularity and volume. In addition, decreases the percentage of DPTCRαβ-/hi cells and increases the percentages of cells within DNTCRαβhi and both SP subpopulations, but much more of the CD8+TCRαβhi subset. These results suggest that the effects of SRIH-14 on the thymus and thymocytes subpopulations are age-dependent.Sa obzirom da literaturni podaci ukazuju da somatostatin ima uticaja na funkciju timusa, kao i da se tokom postnatalnog života menja nivo somatostatina i ekspresija njegovih receptora u timusu, cilj ovog rada je bio da se ispita da li SRIH- 14 davan intracerebroventrikularno u prepubertalnih i mladih odraslih pacova, dovodi do različitih promena u zonama timusa i profilu timocitnih subsetova. Mužjacima pacova AO soja, starim 4 i 10 nedelja, ugrađene su kanile u treću moždanu komoru i oni su tretirani somatostatinom, ukupno tri doze, ubrizgane svakog drugog dana. U prepubertalnih pacova tretman somatostatinom dovodi do smanjenja relativne mase i zapremine timusa, kao i zapremine timusnih zona, naročito dubokog korteksa, što je bila posledica gubitka timocita apoptozom. Takođe, SRIH-14 je doveo do povećanja procenta timocita nezrelog fenotipa (DNTCRαβ-/low, DPTCRαβ-, SPCD8TCRαβ-/low i SPCD4TCRαβ-/low) prekusora DPTCRαβlow subpopulacije, smanjenja procenta DPTCRαβlow i DPTCRαβhi subsetova, dok je relativni odnos najzrelijih CD4TCRαβhi i CD8TCRαβhi timocita ostao neizmenjen. U mladih adulta primena somatostatina, iako dovodi do smanjena zapremine i celularosti korteksa ne dovodi do promena u relativnoj masi timusa. Procenat DPTCRαβ-/hi timocita je bio smanjen, dok je procentualno učešće DNTCRαβhi timocita i obe jednostruko pozitivne subpopulacije (CD4TCRαβhi i CD8TCRαβhi, više CD8TCRαβhi) u ukupnom broju timocita bilo povećano. Navedeni rezultati ukazuju da SRIH-14 različito utiče na morfologiju timusa i na subpopulacije timocita pacova u zavisnosti od uzrasta tretiranih životinja

Absent 99mTc-MIBI Uptake in the Thyroid Gland during Early Phase of Parathyroid Scintigraphy in Patients with Primary and Secondary Hyperparathyroidism

BACKGROUND: Thyroid uptake of technetium-99m methoxyisobutylisonitrile (99mTc-MIBI) during parathyroid scintigraphy can be affected by various conditions.AIM: To evaluate the frequency of absent 99mTc-MIBI uptake by the thyroid gland in the early phase of dual-phase parathyroid scintigraphy.METHODS: The early planar images of dual phase Tc99m MIBI parathyroid scintigraphy from 217 patients performed between 2014 and 2017 were retrospectively analysed. Patients were divided into two groups. The first group included 147 patients with primary hyperparathyroidism and the second group included 70 patients with chronic renal failure. Patient records, laboratory and ultrasonographic data were analysed in all patients. Descriptive statistic was used for data analysis.RESULTS: Out of all patients in the first group, 18 patients (12.24%) showed absent thyroid uptake. Thyroidectomy was performed in 44.4% of these patients, and the rest of them had some thyroid disease. Only one patient had no thyroid or another chronic disease. In the second group, 8 patients (11.42%) presented with absent thyroid uptake of MIBI. Among them, 5 patients had no history of thyroid disease and had been on hemodialysis programme, and 3 patients had hypothyroidism.CONCLUSION: Absent 99mTc-MIBI uptake in the thyroid during the early phase of parathyroid scintigraphy is most frequently related to thyroid disease. A small proportion of patients with chronic renal failure can present with absent 99mTc-MIBI uptake in the thyroid as well. The mechanism for this alteration is still unclear and needs further investigation

Compassionate mesenchymal stem cell treatment in a severe COVID-19 patient: a case report

COVID-19 presentations range from cold-like symptoms to severe symptoms with the development of acute respiratory distress syndrome (ARDS). We report on a severe COVID-19 patient who was mechanically ventilated and who developed ARDS and bacterial infection. Because of rapid clinical deterioration and the exhaustion of other treatment options, the family and attending physicians requested a compassionate use of adult allogeneic bone marrow-derived mesenchymal stem cells (MSC) in addition to commonly used immunosuppressive, antiviral, and supportive therapy. The clinical course is discussed thoroughly, with a special emphasis on the safety and effect of MSC therapy. Compassionate MSC treatment, given in three rounds, affected ARDS regression. The patient was discharged from the intensive care unit after 31 days and from hospital after 49 days in a good general condition. MSC treatment was not associated with any side effects and was well tolerated in a three-week period; therefore, it should be studied in larger trials and considered for compassionate us

Cell proliferation assay - Method optimisation for in vivo labeling of DNA in the rat forestomach

The study of cell proliferation is a useful tool in the fields of toxicology, pathophysiology and pharmacology. Cell proliferation and its degree can be evaluated using 5-bromo-2'deoxyuridine which is incorporated into the newly synthesized DNA. The aim of this study was the optimization of subcutaneous application of 5-bromo-2'-deoxyuridine implantation for continuous and persistent marking of proliferating cells in the rat forestomach. 3-tert-Butyl-4-hydroxyanisole was used as the agent that ensures cell proliferation. In order to determine the optimal dose for proliferating cells labeling, 5-bromo-2'-deoxyuridine doses of 50 mg, 100 mg, 200 mg or 350 mg were implemented 2 days prior to sacrifice by flat-faced cylindrical matrices. Immunohistochemical analysis using 5-bromo-2'-deoxyuridine in situ detection kit was performed for the detection of 5-bromo-2'-deoxyuridine labeled cells. The results showed that for adult rats, the optimum 5-bromo-2'-deoxyuridine dose is 200 mg per animal for subcutaneous application. The here described manner of 5-bromo-2'-deoxyuridine in vivo labeling provides a simple, efficient, and reliable method for cell labeling, and at the same minimizes stress to animals

Cross-Cultural Validation of the Definition of Multimorbidity in the Bulgarian Language.

INTRODUCTION: Multimorbidity is a health issue with growing importance. During the last few decades the populations of most countries in the world have been ageing rapidly. Bulgaria is affected by the issue because of the high prevalence of ageing population in the country with multiple chronic conditions. The AIM of the present study was to validate the translated definition of multimorbidity from English into the Bulgarian language. MATERIALS AND METHODS: The present study is part of an international project involving 8 national groups. We performed a forward and backward translation of the original English definition of multimorbidity using a Delphi consensus procedure. RESULTS: The physicians involved accepted the definition with a high percentage of agreement in the first round. The backward translation was accepted by the scientific committee using the Nominal group technique. DISCUSSION: Some of the GPs provided comments on the linguistic expressions which arose in order to improve understanding in Bulgarian. The remarks were not relevant to the content. The conclusion of the discussion, using a meta-ethnographic approach, was that the differences were acceptable and no further changes were required. CONCLUSIONS: A native version of the published English multimorbidity definition has been finalized. This definition is a prerequisite for better management of multimorbidity by clinicians, researchers and policy makers